| Field | Specification |
|---|---|
| Mfr No | |
| Species |
The Wilms1 cell line was derived from a primary Wilms tumor sample obtained from a patient presenting with large bilateral kidney tumors, indicative of Wilms tumor, a pediatric nephroblastoma. This cell line harbors a homozygous nonsense mutation in the WT1 gene (c.149 C>A, p.S50X), which results in a truncated and non-functional WT1 protein. The WT1 gene, critical for kidney development and function, is frequently mutated in Wilms tumor, particularly in those with a stromal subtype that exhibits ectopic mesenchymal differentiation. Wilms1 cells, therefore, represent a unique in vitro model for studying the consequences of WT1 loss of function in tumor biology.
The Wilms1 cell line maintains a stable karyotype with no significant chromosomal abnormalities, allowing for reliable long-term culture. These cells exhibit a mesenchymal phenotype, characterized by the expression of vimentin and the absence of epithelial markers such as cytokeratin, consistent with their stromal origin. Additionally, the cell line demonstrates limited but notable mesenchymal differentiation capacity, including the ability to differentiate into muscle-like cells under appropriate conditions. This makes Wilms1 an invaluable tool for investigating the molecular mechanisms of mesenchymal differentiation and its deregulation in Wilms tumor pathogenesis.
Wilms1 has also been used to study the activation status of key signaling pathways involved in tumor progression. Proteomic analyses have shown that Wilms1 cells exhibit phosphorylation and activation of several receptor tyrosine kinases, including EGFR and PDGFRβ, as well as downstream MAPK signaling pathways. These findings highlight the relevance of the Wilms1 cell line in exploring targeted therapeutic approaches for Wilms tumor by dissecting the role of these pathways in cancer cell survival, proliferation, and differentiation.
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- Receptors expressed: Receptor tyrosine kinases EGFR, EphA7, PDGFRalpha, FGFR1, PDGFRbeta, AxL
- Tumorigenic: Yes, in nude mice. Forms tumor with small cells consistent with Wilms' tumor (xenografts may not represent Wilm’s tumors completely, see E. Kunce Stroup 2017)
- Viruses: HIV-1: negative, HBV: negative, HCV: negative
- Mutational profile: WT1 mutation status: homozygous c. 149 C>A, p.S50x, LOH: 11p11-11pter, CTNNB1 mutation status: heterozygous TCT>TTT, p.S45F
- Karyotype: 46, normal
- cultureMedium: MSCGM kit (from Lonza)
- dissociationReagent: Accutase
- doublingTime: 24 hours
- subculturing: Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium.
- seedingDensity: 1 x 104 cells/cm2
- fluidRenewal: 1 to 2 times per week
- postThawRecovery: Fast
- freezeMedium: As a cryopreservation medium, use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress.
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